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Method, probe and kit for detecting P16 cancer suppressor gene deletion

A tumor suppressor gene and gene probe technology, applied in the field of malignant tumor-related gene detection, can solve problems such as inaccurate results, and achieve the effects of simple operation, comprehensive design, and accurate and reliable detection results.

Inactive Publication Date: 2019-03-29
江苏蒙特立生物技术有限公司
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Problems solved by technology

[0004] At present, PCR method is mainly used to detect P16 gene deletion in the market. Since the detection is a mixture of tumor and normal cells, the result is not accurate enough, and a more accurate detection method is urgently needed in the market.

Method used

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  • Method, probe and kit for detecting P16 cancer suppressor gene deletion

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Embodiment Construction

[0013] In order to better understand the content of the present invention, the following will be further described in conjunction with specific examples. The main reagent that following specific embodiment adopts, instrument and operating procedure are as follows:

[0014] 1. Acquisition of probe DNA

[0015] 1) Streak inoculation: Streak inoculate the BAC strain carrying the P16 gene and the centriole of chromosome 9 on an agar plate containing an antibiotic (chloramphenicol), and culture at 37° C. overnight (about 14 hours).

[0016] 2) Initial culture: Pick a monoclonal strain with good growth status the next day, inoculate it in 2ml of LB liquid medium containing antibiotics (chloramphenicol), put it in a shaker at 37°C, set the shaker speed to about 200rpm, and culture 6-8 hours.

[0017] 3) Overnight culture: transfer each 1ml of bacterial liquid to 200ml culture system, place on a shaker at 37°C with a rotation speed of 200rpm, and culture overnight (about 14-16 hours...

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Abstract

The invention relates to a method, a probe and a kit for detecting P16 cancer suppressor gene deletion. The method includes: a group of two-color probes are used for in-situ hybridization on cancer tissue, and according to detected fluorescent signal, existence of the P16 cancer suppressor gene deletion is determined. The group of two-color probes includes: 1) a P16 gene probe labeled as red and 2) a #9 chromosome centriole probe labeled as green. The method is comprehensive and is simple in operation, has high specificity and sensitivity, can quickly, accurately and directly detect the deletion of the P16 cancer suppressor gene, is accurate and reliable in detection result, and can be used as a reference for molecular pathological typing on malignant tumors by doctors, and is suitable forlarge-scale promotion and application.

Description

technical field [0001] The invention relates to the technical field of detection of malignant tumor-related genes, in particular to a detection method for the deletion of a P16 tumor suppressor gene, a related detection probe and a kit. Background technique [0002] The p16 gene, also known as MTS (multiple tumor suppressor 1) gene, is a new anti-cancer gene discovered by Kamb of Cold Spring Harbor Laboratory in the United States in 1994. It is a basic gene in the cell cycle, directly involved in the regulation of the cell cycle, negative regulation Cell proliferation and division. Homozygous deletion and mutation are found in 50% of human tumor cell lines. It is believed that p16 is a new type of anti-cancer gene that is more important than p53. Some people compare it to the brake device in the cell cycle. Once it fails It will lead to malignant proliferation of cells, leading to malignant tumors. P16 gene has been found in lung cancer, breast cancer, brain tumor, bone tum...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886
CPCC12Q1/6841C12Q2563/107
Inventor 缪为民
Owner 江苏蒙特立生物技术有限公司
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